【關鍵詞】 心肌上清液
Apoptosis of mouse myeloma Sp2/0 cells induced by supernatant of mouse cardiac muscle lysate
【Abstract】 AIM: To study the roles of mouse cardiac muscle lysate supernatant in inducing the apoptosis of mouse myeloma Sp2/0 cells. METHODS: Various concentrations (1∶6, 1∶12, 1∶24) of mouse cardiac muscle lysate supernatant and cyclophosphamide were added into the Sp2/0 cell culture wells. After 24 h, microscope was used to observe the morphological changes of Sp2/0 cells and apoptosis rate was determined by flow cytometry after 48 h. Morphological changes of Sp2/0 cells cocultured with mouse cardiac muscle cells were observed after 10 h. The apoptosis of Sp2/0, Hela, Hep2 and Vero cells was induced with various concentrations (1∶5, 1∶10, 1∶20 and 1∶40) of mouse cardiac muscle lysate supernatant, liver lysate supernatant, thymus lysate supernatant and cyclophosphamide, and then the difference of apoptotic rate among them was compared. RESULTS: Mouse cardiac muscle lysate supernatant could induce the apoptosis of the Sp2/0 cells and the apoptotic rate increased in a concentrationdependent manner, but there was no difference between mouse cardiac muscle lysate supernatant group and cyclophosphamide group. After a 10hour coculture with mouse cardiac muscle cell lysate, the apoptosis of a few Sp2/0 cells was seen. The mouse cardiac muscle lysate supernatant, liver lysate supernatant and thymus lysate supernatant could all induce apoptosis of the Sp2/0, HeLa, Hep2 and Vero cells. But the effect of the mouse cardiac muscle lysate supernatant was more obvious and permanent. CONCLUSION: The mouse cardiac muscle lysate supernatant can significantly induce apoptosis of mouse myeloma Sp2/0 cells.
【Keywords】 cardiac muscle lysate supernatant; apoptosis; mouse myeloma Sp2/0 cell
【摘要】 目的: 研究小鼠心肌上清液誘導小鼠骨髓瘤Sp2/0細胞凋亡的作用. 方法： 在Sp2/0細胞培養孔中，分別加入不同稀釋度(1∶6, 1∶12, 1∶24)的小鼠心肌上清液和環磷醯胺， 24 h後鏡下觀察細胞形態，並於48 h後流式細胞儀檢測凋亡細胞的比率；同時，Sp2/0細胞和小鼠心肌細胞共培養，10 h後觀察Sp2/0細胞形態學變化；並用不同稀釋度（1∶5, 1∶10, 1∶20, 1∶40）小鼠心肌裂解上清液、肝臟裂解上清液、胸腺裂解上清液及環磷醯胺誘導Sp2/0, HeLa, Hep2和Vero細胞凋亡，並比較其差異. 結果: 不同稀釋度的`心肌裂解上清液均可誘導Sp2/0細胞的凋亡，尤以高稀釋度更明顯；小鼠心肌細胞與Sp2/0細胞共培養10 h後,少部分Sp2/0細胞出現凋亡；小鼠心肌裂解上清液、肝臟裂解上清液、胸腺裂解上清液均可引起Sp2/0, HeLa, Hep2和Vero細胞發生凋亡，但小鼠心肌裂解上清液作用明顯，且作用時間持久. 結論: 小鼠心肌組織裂解上清液具有明顯誘導小鼠骨髓瘤Sp2/0細胞凋亡的作用.
【關鍵詞】 心肌上清液；細胞凋亡；小鼠骨髓瘤Sp2/0細胞
0引言
細胞凋亡與腫瘤的發生發展密切相關，已成為研究熱點［1］，許多預防治療腫瘤的藥物、細胞因子等抑制腫瘤細胞生長的機制之一是誘導腫瘤細胞凋亡. 我們從細胞凋亡的角度探討心肌上清液誘導小鼠骨髓瘤Sp2/0細胞凋亡.
1材料和方法
1.1材料
Balb/c 20只8周齡小鼠，雌雄各半，體質量約250 g，由安徽安科生物工程股份有限公司動物室提供；環磷醯胺(2 g/L)由湖北科利藥業股份有限公司生產; DMEM液由美國生命技術公司生產；胎牛血清由杭州四季青公司提供；流式細胞儀（日本產）；Leica熒光顯微鏡(德國Leica公司)；CO2溫箱(美國產)；紫外分光光度計（上海產）. Sp2/0, HeLa, Hep2和Vero細胞株均來源於中國預防科學院病毒所，均以100 mL/L胎牛血清＋DMEM液為培養液，按本室常規方法在CO2溫箱中培養；乳鼠心肌細胞培養： 取Balb/c新生紅皮乳鼠20只，在無菌操作下取出心臟，用PBS沖洗3遍，以眼科剪剪成1 mm3碎塊，洗滌，後加2.5 g/L胰蛋白酶液37℃消化20 min，吹打3次，吸取上層細胞懸液，以同樣的方法消化3次，直到組織塊完全消化為止，以1500 r/m